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PubMed1994

Mycoplasma and Chlamydia in adenoids and tonsils of children undergoing adenoidectomy or tonsillectomy.

The prevalence of mycoplasmal and chlamydial infection was assessed in 83 children undergoing adenoidectomy, tonsillectomy, or both procedures for recurrent adenotonsillitis or obstructive symptoms. Throat smears (surface specimens) and minced adenoids and tonsils (core specimens) were cultured for Mycoplasma spp and for Chlamydia spp. Isolation rates in adenoidal specimens were as follows: Mycoplasma hominis, surface 7.1% core 2.9%; and Ureaplasma urealyticum, surface 1.4%, core 2.9%. Mycoplasma hominis was also found in tonsillar specimens: surface 14.3%, core 20%. Chlamydia trachomatis was isolated only from a single core adenoidal specimen. The rate of mycoplasma isolation was significantly higher in children with recurrent adenotonsillitis (34.5%) than in those with obstructive symptoms (3.7%). Our findings document colonization of genital mycoplasmas in adenoids and tonsils of children with recurrent adenotonsillitis. Further studies are needed to evaluate the possible pathogenetic role of these microorganisms in adenotonsillar infection.

Adenoidectomy↗

PubMed1976

A highly infectious 'mycoplasma' that inhibits meiosis in the fungus Coprinus.

We have discovered a cytoplasmically inherited infectious agent that inhibits meiosis in a species of Coprinus, a basidiomycetous fungus. From infectivity, filtration, centrifugation and ultrastructural studies we believe the agent to be a mycoplasma. The agent is highly infectious to several strains of the host species and is capable of spreading rapidly through infected hosts. No pathological effect has been seen on any aspect of growth or differentiation of the fungus except for the inability of infected strains or undergo meiosis. The failure of meiosis results in mushrooms that do not produce the normal black spores and are therefore pale in colour. The paleness represents a simple assay for the presence and activity of the infectious agent. Infected hosts do not display any ultrastructural abnormalities in the vegetative stages, only in the cells in which meiosis should occur. In the meiotic cells, at the time when normal cells are undergoing synapsis and synaptinemal complexes are forming, the vacuoles of the infected cells become occupied with vesicular, membrane-bound bodies resembling in shape and form mycoplasmas. Extracts from infected clones may be filtered through 0.2-mum filters and retain full infectivity. The infectious material may be pelleted from such extracts at only 10 000g. Migration experiments, as well as the filtration studies, rule out involvement directly of nuclei. The high rate of infection and spread of the mycoplasma through the host, combined with the anatomical simplicity of the host, make this an ideal system in which to study the basis of infection. The singularity of the pathological effect make this host-parasite association useful in studying both the underlying mechanisms of mycoplasma pathogenicity and to investigate the regulation of meiosis. This is only the second report of mycoplasmas in fungi.

Agaricales↗

PubMed1997

Survival of mycoplasmas inoculated in horse sera.

Although it is known that commercialized bovine serum is sometimes contaminated with mycoplasmas, it is not clear whether mycoplasmas can survive in horse serum. In this study, as a preliminary examination of the survival of mycoplasmas inoculated in horse sera, the survivability of 8 strains of 7 mycoplasmas was tested. The results obtained reveal that two strains of M. bovis and M. gallisepticum were found to survive in non-heated and inactivated sera for 94 to 330 days at 30 or 37 degrees C. Three strains of M. bovirhinis, M. gateae and A. laidlawii lived for 7 to 330 days depending upon the temperature maintained or pH of the serum. Strains of M. bovigenitalium and U. diversum survived for a maximum of 8 days in all horse sera tested. Therefore, mycoplasmas are generally likely to survive for a long period in horse serum although the survival period depends on the species, strain and temperature.

Animals↗

PubMed1982

Effect of Mycoplasma pulmonis on in-vivo fertilization in the mouse.

When fresh, epididymal mouse spermatozoa were incubated with viable Mycoplasma pulmonis, the mycoplasma adhered to the heads and tails of spermatozoa. Spermatozoa that had been preincubated with M. pulmonis in phosphate-buffered saline (PBS) and spermatozoa incubated in PBS alone were inseminated into the uterine horns of mice induced to superovulate. The mycoplasma treatment resulted in a reduced rate of fertilization and a decrease in the number of spermatozoa associated with eggs recovered from the oviduct at 5--18 h after ovulation. The percentage of spermatozoa recovered from the oviduct with adherent mycoplasmas, as determined by fluorescence microscopy, was lower than that seen in the uterus. Mycoplasma treatment of spermatozoa inseminated directly into the ovarian bursa did not result in a reduced fertilization rate. The results suggest that M. pulmonis adversely affects sperm transport through the female reproductive tract in the mouse.

Animals↗

PubMed2000

Disease caused by Mycoplasma mycoides subspecies mycoides LC in Hungarian goat herds.

The occurrence of a goat disease caused by Mycoplasma mycoides subsp. mycoides LC in Hungary is reported. The disease occurred in two goat herds in the spring of 1999. In one herd 25% of the 4-12 weeks old kids (10 animals) while in the other herd 33% of the 6-12 weeks old kids (20 animals) became affected. The goat kids developed polyarthritis. The most severe lesions developed in the carpal joints. All animals died after 3-8 days of disease. Four dead kids were necropsied. All of them had serofibrinous and purulent polyarthritis, and in two animals bronchopneumonia, fibrinous pleuritis and meningitis were also found. In the articular exudates the presence of mycoplasmas was detected by PCR using a general mycoplasma primer. Mycoplasmas were cultured from the joints of all animals, from the abdominal parenchymal organs of two kids and from the lungs of one animal. The cultured mycoplasmas grew in strikingly large colonies, proved to be glucose positive, arginine negative and phosphatase positive, and liquefied the coagulated serum. They survived incubation at 45 degrees C for more than 24 h. Based upon their biochemical properties, the results of the immunofluorescence (IF) and growth inhibition tests and the sequence analysis of the PCR product, the cultured strains were identified as M. mycoides subsp. mycoides LC. Animals purchased in the previous autumn had been introduced to both farms. The disease may have been introduced with asymptomatic carrier animals, as earlier no similar disease had been observed at either farm.

Animals↗

PubMed1989

[Mycoplasma as a contaminant of cell cultures maintained in laboratories of private and official institutions].

Mycoplasma is one of the most serious contaminants of cell cultures. Its detection is very important in virology, as well as its eradication. The aim of this study was to verify the incidence of mycoplasma in cell lines maintained in seven laboratories of private, government and college institutions of the State of São Paulo, Brazil, for the purposes of research, production of reagents for diagnosis and production of biologicals for human and animal use. Of the 29 cell lines, eight were derived from human tissues and 21 from other animal species (dog, rabbit, mouse, hamster, monkey, pig, chicken and ox). Using the direct method with specific liquid and solid media for detection of mycoplasma, 48 out of the 106 cell samples tested were positive, corresponding to a contamination index of 45.28%. The incidence of contamination among the 35 cell samples of human origin was 51.43% (18 positive). Of the 71 samples originated from other species, 30 were positive (42.25%). The high incidence of contamination found calls for the adoption of measures for the prevention of this hazard: the elimination of mouth pipetting, the use of aseptic techniques and a rigid control of trypsin, serum and other components of cell culture media. The substitution of mycoplasma-free cultures for all contaminated ones and the performance of periodical tests for mycoplasma detection must also be carried out to prevent and avoid the dissemination of these organisms. Data obtained showed that contamination appeared in the 2nd (72.92%), in the 3rd (20.83%) and in the 4th passage (6.25%).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1995

Mycoplasmas in plaque and saliva of children and their relationship to gingivitis.

Gingivitis is common in children but its etiology is not well understood. Results of earlier studies have suggested that mycoplasmas may have a role in the disease. In this study two plaque samples and one stimulated saliva sample were taken from each of a group of sixty (60) 5 to 9 year-old children and cultured selectively for mycoplasmas. Subjects had been selected on the basis of gingival status buccal to the most posterior tooth in an upper quadrant, 20 each having scores 0, 1, or 2 at this site using the Löe and Silness index. Plaque samples were taken from the adjacent tooth surface and from a second posterior site where the adjacent gingiva was free of signs of inflammation. Mycoplasmas were cultured from 54 saliva samples and from 57 plaque samples. Isolation frequency and numbers of mycoplasmas in relation to total anaerobic counts were consistently related to gingivitis scores in both plaque and saliva samples, with higher frequencies and proportions with increasing levels of gingival inflammation at the selected site. In plaque samples this finding held good for both comparisons between groups of subjects and for those between test and control sites within subjects. Findings would appear to confirm the suggestions of earlier studies that mycoplasmas in the oral cavity are associated with gingivitis in children.

Chi-Square Distribution↗

PubMed1997

Detection of mycoplasma infection of mammalian cells.

Mycoplasma infection was detected in cultures of COS cells with a novel, simple assay that detects the conversion of arginine to citrulline by the enzyme, arginine deiminase, specific to all species of mycoplasma. Transfection of COS cells was inhibited in mycoplasma-infected cells, a phenomenon that was readily reversed by treatment with a mycoplasma removal agent. Cultures of cells used for transfection should be regularly monitored for evidence of mycoplasma by assay of arginine deiminase activity or by other methods.

Animals↗

PubMed1984

The infrequent occurrence of mycoplasmas in amniotic fluid from women with intact fetal membranes.

As human genital mycoplasmas have been associated with various forms of reproductive failure, the present study was undertaken to investigate whether M. hominis and U. urealyticum organisms (ureaplasmas) are capable of crossing intact fetal membranes. Nearly 300 women in Denmark and England were investigated. Most of them were seen at about the fourth month of gestation and the remainder towards or at the time of birth, all with unruptured membranes. A swab was taken from the uterine cervix or vagina and M. hominis was isolated from 9% of the women and ureaplasmas from half of them. The presence of these mycoplasmas was not associated with an abnormal outcome of pregnancy. In contrast to the frequent presence of mycoplasmas in the lower genital tract, amniotic fluids obtained by transabdominal amniocentesis or at cesarean section did not contain M. hominis and ureaplasmas were isolated from only one of them. This was associated with the same ureaplasmas serotype being recovered from the cervix and also from the blood of both infant and mother, whose case differed from the others as labor had already started when the amniotic fluid was obtained. Thus, in our populations, we have no evidence that mycoplasmal invasion of the amniotic fluid occurs before the onset of labor. During labor, despite intact membranes, it seems that genital mycoplasmas may occasionally invade the fetal--placental unit, probably by the hematogenous route after strong uterine contractions, or otherwise directly after membrane rupture. Since both these events are followed usually by immediate delivery, there would seem to be insufficient time for the genital mycoplasmas to cause fetal damage.

Amniotic Fluid↗

PubMed2004

Effects of storage time and thawing methods on the recovery of Mycoplasma species in milk samples from cows with intramammary infections.

This study was executed to determine the effects of storage and thawing on the viability of Mycoplasma spp. in milk from cows with intramammary infections. The trial was designed using a control sample and seven handling regimens subjected to two methods of thawing. There was a significant treatment effect on the recovery of colony-forming units in milk samples when comparing the control sample with handling regimens 1 through 7. There was a continuous decline in log (10) mean number of cfu/mL recovered. Mean concentrations were 6.29, 4.64, 3.69, 3.01, 1.86, 4.41, 4.13, and 3.18 for control and handling regimens 1 to 7, respectively. To determine the best thawing method, handling regimen 1 through 7 samples were thawed using two methods. On average, more mycoplasma were recovered from milk samples thawed at ambient temperature (4.04 cfu/mL) than milk samples thawed in a 37 degrees C water bath (3.76 cfu/mL). A final comparison was made between individual treatments. With the exception of the handling regimen 5 to 6 pair-wise comparison, all pair-wise comparisons between handling regimens were significantly different. The results of this study indicate that storage and thawing of milk samples is harmful to mycoplasma organisms. Fresh samples should be used to improve detection of Mycoplasma spp. from milk of infected cattle. If frozen samples are used, then length of storage time should be minimized, and thawing milk at ambient temperature will improve recovery of mycoplasma as opposed to thawing in a 37 degrees C water bath.

Animals↗

PubMed1978

[Identification of various porcine mycoplasmas using epi-immunofluorescence].

The responses of the field strains and type strains of Mycoplasma hyorhinis are compared in epi-immunofluorescence, in growth-inhibition tests, and growth-precipitation tests. The type strains, together with the field strains, could be definitely identified by direct and indirect epi-immunofluorescence. In indirect epi-immunofluorescence, all rabbit antiserums against the strains of Mycoplasma hyorhinis tested by us always gave positive cross reactions with all the tested strains of Mycoplasma hyorhinis. On the other hand, in the growth-inhibition cross test and growth precipitation cross test, the type strains of Mycoplasma hyorhinis always reacted positively only with the homologous antiserums, whereas the heterologous reactions were mostly negative or dubious. The clearly positive epi-immunofluorescent reactions with serums against any strain of Mycoplasma hyorhinis suggest that this method is more suitable for the identification of Myocplasma species than the other two tests studied.

Animals↗

PubMed1999

Detection of mycoplasma contaminations in cell cultures by PCR analysis.

Mycoplasma contamination is still one of the main problems in using cell cultures in biological and medical research and in the production of bioactive substances, because mycoplasma can alter nearly all parameters and products of the cell. They can persist undetected in the culture if no special detection methods are applied. In recent years, the PCR technology has become a commonly used method to analyze genomic DNA and the expression of genes, with both high specificity and sensitivity. This technique can be effectively employed for the detection and even the identification of mycoplasma contaminations in cell cultures applying primers complementary to the 16S rDNA region. Although this technique, once established, is characterized by simplicity and speed, PCR is still a complex process and its sensitivity and specificity can be influenced by a number of different parameters, e.g. inhibiting compounds originating from the preparation process of the DNA, RNA or cDNA, contamination of the solutions with PCR products, and the selection of a primer pair which does not cover all the mycoplasma species occurring in cell cultures. Thus, adequate controls have to be included to obtain reliable results. The present review examines the use of different primers of the 16S rDNA region including their specificity, the sensitivity applying various DNA or RNA preparation procedures, and the methods to detect finally the amplicons. In conclusion, basic nucleic acid preparation and PCR product detection methods offer a simple, fast and reliable technique for the examination of mycoplasma contaminations in cell cultures, provided that the indispensable control assays are implemented.

Animals↗

PubMed2001

[Interaction of Mycoplasma with immune system of animals and humans].

Mycoplasmal infections of animals and human belong to latent or chronic ones, being commonly accompanied with immunodeficiency symptoms. The following topics are discussed in this review: 1) a direct influence of mycoplasmas on the immune system cells with an interference and compression of the functional activity of these; 2) influence of mycoplasmas on the immune system via cytokine induction; 3) mitogenes and mitogenic action of mycoplasmas; 4) the role of mycoplasmas in the development of autoimmune processes. Basic host immune response to mycoplasma is presented in diagrams.

Animals↗

PubMed2001

Serum interleukin-6 and tumor necrosis factor-alpha concentrations in children with mycoplasma pneumonia.

A prospective study was performed to assess the relationship among interleukin-6, tumor necrosis factor-alpha, C-reactive protein serum concentrations, and the severity of mycoplasma pneumonia in 49 children. Mycoplasma pneumonia was diagnosed by chest film and anti-Mycoplasma pneumoniae IgM antibody test. Serum concentrations of interleukin-6 and tumor necrosis factor-alpha were measured by using enzyme-linked immunosorbent assays. Interleukin-6 serum levels in mycoplasma pneumonia patients with fever for more than 3 days (41.98 +/- 67.46 [SD] pg/mL) were significantly higher than those in patients with fever < or = 3 days (10.01 +/- 11.74 pg/mL, p < 0.05). Interleukin-6 serum levels in those patients whose chest films revealed patchy consolidations or pleural effusion (58.11 +/- 92.19 pg/mL) were significantly higher than those in patients whose chest films revealed peribronchial interstitial infiltration (15.94 +/- 20.81 pg/mL, p < 0.05). The mean levels of serum tumor necrosis factor-alpha were not statistically significant in the different duration of fever and chest film findings. These results suggest that interleukin-6 serum concentration, rather than tumor necrosis factor-alpha, may be a potential indicator of the severity and outcome of mycoplasma pneumonia.

C-Reactive Protein↗

PubMed2001

[Pneumonia due to mycoplasma pneumoniae in a primary health care district].

OBJECTIVE: Most studies of atypical pneumonia due to Mycoplasma pneumoniae have been performed in hospitalized patients. The aim of this study was to determine the epidemiological, clinical and radiological characteristics as well as the evolution of patients with atypical pneumonia due to Mycoplasma pneumoniae in an out-of-hospital setting. METHODS: A prospective observational study was conducted in 31 patients with atypical pneumonia due to Mycoplasma pneumoniae in the pediatric population of a primary health care district from May to July 1996. In all patients serological confirmation of infection by Mycoplasma pneumoniae was obtained using a complement-fixation test. RESULTS: The age of the children in the study ranged from 4-13 years. The most common clinical manifestations were cough (93 %), high fever (84 %) and pharyngitis (48 %). The most frequent auscultatory finding was crackles (93 %), which were bilateral in 14 patients. Clinical-radiological dissociation was found in 16 % of the children; 27 (87 %) showed radiological alterations. No characteristic radiological pattern was detected because alveolar and interstitial alterations were equally frequent, with a predominance of lung base involvement (67.7 %). In all patients response to macrolides was excellent, fever abated within 48-72 h and the remaining symptoms progressively improved. CONCLUSIONS: Atypical pneumonia due to Mycoplasma pneumoniae normally presents in children aged more than 5 years old or in adolescents. No signs, symptoms, or radiological patterns clearly indicate the etiology. However, because of its frequency, this disease should be suspected when school-aged children or adolescents present a pneumonia syndrome. Macrolides administration

Child↗

PubMed2001

[Mycoplasma pneumonia complicated by acute renal failure].

A 17-year-old woman presented with cough and fever. Treatment with cefteram pivoxil and fosfomycin calcium was not effective. Chest radiography showed infiltration with an air bronchogram in the left upper lung field. The case was initially treated as atypical pneumonia by administering minocycline hydrochloride and piperacillin sodium. Since acute renal failure progressed rapidly, we introduced hemodialysis, and renal function improved. The mycoplasma polymerase chain reaction (PCR) in the pharynx smear was positive, and the mycoplasma serum titer was 1280 on admission. On the basis of these findings, mycoplasma pneumonia was diagnosed. Only one other case of mycoplasma pneumonia with hemodialysis has been reported in Japan. We report and assess the pathogenesis of mycoplasma pneumonia complicated by renal dysfunction.

Acute Kidney Injury↗

PubMed1976

The role of Mycoplasma in human infertility.

Seminal fluid from males complaining of infertility was examined for the presence of T-mycoplasmas and Mycoplasma hominis. No significant difference, with regard to the presence of mycoplasmas, was found between the seminal fluid which had normal cytology and seminal fluid of which the cytology was deemed pathological. Colonisation was shown to be an evanescent phenomenon, and female consorts often had different mycoplasma flora to the males. No positive role of mycoplasmas in the pathogenesis of infertility could be shown.

Cervix Uteri↗

PubMed1976

[Differentiation of swine-Mycoplasma by latex-agglutination (author's transl)].

UNLABELLED: The strains of Mycoplasma were grown in Shittlestone medium for 3-5 days. After centrifugation and washing, the mycoplasm were freeze-dried and stored at -20 degrees C. From this stock material immune sera and antigens were prepared. The immune sera were prepared by immunization of rabbits with a suspension of mycoplasma (2,2 mg dry weight per ml) and complete Freund's adjuvant. For the first immunication, 6.6 mg of antigen were injected into rabbits at different sites. 3 weeks later the second immunization followed by the intramuscular route (4.4 mg of antigen). The third and fourth immunizations were identical to the second one. Antiserum was obtained as usual. In order to avoid unspecific reactions in the latex agglutination, 1 ml of antiserum was added for absorption to 200 mg of freeze-dried sterile Whittlestone medium over 2 days at 4 degrees C. Antigen was prepared by ultrasonic disruption of mycoplasma (20 mg dry weight/ml physiological saline) and centrifugation 17300 X g, 30 min). The supernatant was used as antigen. The latex agglutination was performed in 0.1 M borate buffer pH 8,2. For preparation of the latex antigen compound 0.2 ml of a latex suspension (diluted 5 fold with water) were moxed with 0.5 ml of an antigen suspension (diluted 16 fold with buffer). After 10 min at room temperature, 4.3 ml of borate buffer were added. After 10 min, 0.3 ml of this solution were added again each to 0.3 ml of serial dilutions of anitserum. The suspensions were mixed and left for 20 h at 37 degrees C and after this time for 1 h at room temperature. Then the reactions were read. RESULTS: M. suipneumoniae and M. hyopneumoniae were found to be identical or very close related strains. Between M. hyorhinis and M. sp. E9 relationship was noted too. M. granularum was different to all other investigated strains of mycoplasma. All results of Latex agglutination are in agreement with investigations performed with the aid of other serological methods and of electrophoretic separations of cell proteins of these mycoplasma.

Animals↗